①用EFS30、EFS40、EDFS30、EDFS40四种玻璃化冷冻液对MⅡ期水牛卵母细胞进行毒性试验,结果表明:试验组卵母细胞形态正常率与对照组均无显著性差异(P＞0.05)；对卵母细胞孤雌激活后EDFS30、EDFS40组的卵裂率与对照组(75.28％)及EFS30、EFS40组差异显著(P＜0.05)；利用4种冷冻保护剂采用OPS法冷冻保存MⅡ期水牛卵母细胞,其中以EDFS40作为冷冻液时,卵母细胞冷冻解冻后孤雌激活卵裂率最高,达31.60％；以EDFS40作为冷冻液,比较了GMP法和OPS法的冷冻效果,结果表明GMP法冷冻效果好于OPS法.②采用不同预处理时间和平衡时间使用细管法常规冷冻GV期卵母细胞,结果表明预处理5 min、平衡15min组的形态正常率和极体排出率相对较高,分别为72.73％、27.27％.%This study was first employed to investigate the developmental potential of in vitro matured buffalo oocytes vitrified by open-pulled straw (OPS) and glass micropipette(GMP) methods. The oocytes were first pretreated in 10 % EG for 0.5 min and then 25 sec equilibration in EFS30 (group 1) or EFS40 (group 2) ,or in 10 %EG+10 %DMSO for 0.5 min and then 25 sec equilibration in EDFS30 (group 3) or EDFS40 (group 4) before they were subsequendy diluted in 0.25 mol/L sucrose for 1 min and 0.15 mol/L sucrose for 5 min. After dilution,the morphologically normal rates of the oocytes in the 4 groups were similar to that of the control. After parthenogenetic activation, the cleavage rate of oocytes in group 3 and group 4 were significantly lower (P<0.05 ) than that in group 1 and group 2, which were similar to that of control (75.28%). After OPS vitrification and wanning, the oocytes in group 4 showed the highest cleavage rate (31.60%). However, when the oocytes in group 4 were cryopreserved by GMP method,the cleavage rate were 45.63%. Secondly, the G V oocytes were cryopresered by slow freezing. After thawing, the morphologically normal rate and the polar body extrusion rate of the oocytes were significantly lower when compared with those of control.
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